利用复合微生物菌剂控制水华的治理工程试验

为了探索利用微生物控制正在爆发水华的富营养化城市内湖的效果,试验利用光合细菌(Rhodop seudomonas palustris)、芽孢杆菌(Bacillus subtilis)2种微生物菌剂,采用直接投放的方式,治理武汉市重度富营养化湖泊四美塘湖(西),治理时间从2008年8月1日持续到2008年9月27日,治理结束后,湖水水质从劣Ⅴ类提高到Ⅴ类,高锰酸钾盐指数除去率达52.78%,TN除去率达49.26%,TP除去率达73.02%,氨氮除去率达16.18%,Chl-a含量降低了46.70%。治理进行1个月后,湖水的感官效果大幅提高,藻类水华完全消失,透明度提高,不良气味消失。在投菌结束后的3个月内,湖水水质仍然保持良好,没有出现反弹。利用复合微生物菌剂治理富营养化湖泊,控制水华的爆发,是一种经济、快速、无二次污染的富营养化水体治理方法,如将其与其他的治理技术相结合将会使其在富营养化水体治理领域中发挥重要的作用。     

苏云金芽胞杆菌YBT-1520中SigL及其增强子结合蛋白的结构与功能分析

为揭示SigL及其增强子结合蛋白(EBPs)在苏云金芽胞杆菌(Bt)中的调控功能,在全基因组测序的基础上,采用生物信息学方法,对YBT-1520菌株的SigL及其EBPs的结构和功能进行深入分析.结果表明,YBT-1520基因组中存在1个SigL和6个EBPs,而且EBPs在结构上具有丰富的多样性,包含了EBPs的所有可能的结构域组织类型.SigL所调控的基因涉及11个假定的COG代谢途径,其中包括能量代谢、氨基酸代谢、翻译与细胞周期等.根据EBPs在基因组的位置推测,YBT-1520的EBPs参与γ-氨基丁酸代谢途径、精氨酸代谢途径、支链脂肪酸降解途径、多糖分解代谢等代谢途径的调控.本研究将为揭示Bt杀虫晶体蛋白大量表达的调控机制提供新的思路.     

对甘薯茎线虫具有杀虫活性的苏云金芽胞杆菌筛选及特性鉴定

为筛选对甘薯茎线虫具有高毒力的苏云金芽胞杆菌(Bacillus thuringiensis,Bt)菌株,提取12株供试Bt菌株的晶体蛋白,溶解后加入到甘薯茎线虫悬浮液中,统计3 d和7 d后甘薯茎线虫的死亡率.经初筛和复筛后得到一株对甘薯茎线虫具有高毒力的Bt菌株YBT-008,其对甘薯茎线虫的LC50值为203.76μg/mL.研究了YBT-008的生物学特性,结果表明,YBT-008在LB培养基上培养约12 h后进入稳定期,并且伴随芽胞的形成产生卵圆形的伴胞晶体;SDS-PAGE分析表明YBT-008可产生多种待鉴定的晶体蛋白类型,质粒检测显示该菌株有5条质粒条带.YBT-008的获得为利用Bt防治甘薯茎线虫提供了新型菌株和基因资源.     

角蛋白酶产生菌的分离鉴定及其kerC的克隆表达

以四川农业大学养鸡场堆砌废弃羽毛处土壤为样品,筛选出一株具有较强降解羽毛能力的细菌B-3菌株.经形态学、生理生化特性和16S rRNA基因序列分析,鉴定其为枯草芽孢杆菌(Bacillus subtilis),命名为枯草芽孢杆菌B-3.并成功克隆到该菌株的角蛋白酶基因kerC(GenBank No.:JN021789),在大肠杆菌BL21(Escherichia coli BL21)中获得了高效表达.该基因全长1146bp,GC含量46.5%,编码381个氨基酸,与已报道的枯草芽孢杆菌YYW-1的kerC基因(GenBank No.: EU362730)同源性达到100%.重组菌株经IPTG诱导后角蛋白酶酶活力达14.8U/mL,经His-Tag纯化和SDS-PAGE分析表明,重组角蛋白酶分子量约为60kDa(融合了硫氧还蛋白,Trx).重组角蛋白酶最适反应温度和pH值分别为65℃与7.0.     

硝基苯好氧降解菌筛选及其降解特性

针对化学试剂厂和制药厂废水中的硝基苯,经菌源筛选与长期好氧驯化,分离到1株能有效降解硝基苯的菌株Bacillus subilis(枯草芽孢杆菌)。系统研究了其生长条件及降解硝基苯的特性,适宜生长条件: pH为5.0～8.0,温度为30～40℃,NH4Cl浓度为100～200 mg/L,在降解硝基苯的过程中有苯胺生成,这为含低浓度硝基苯废水的生物处理提供了新的途径,即不需要厌氧工艺而直接用好氧技术就可将废水完全无害化。      

枯草芽孢杆菌对铜绿微囊藻抑制效果的研究

为探讨枯草芽孢杆菌(Bacillus subtilis)对铜绿微囊藻(Microcystis aeruginosa)的抑制效果,在实验室条件下,研究了枯草芽孢杆菌不同生长时期(延迟期、对数期、稳定期和衰亡期)无菌滤液对铜绿微囊藻生长的影响、枯草芽孢杆菌抑制铜绿微囊藻生长的作用方式以及无菌滤液影响下铜绿微囊藻丙二醛(MDA)含量、超氧化物歧化酶(SOD)活性和光合色素含量的变化.结果显示:枯草芽孢杆菌对数期、稳定期和衰亡期滤液抑藻效果明显好于延迟期,作用第8d,对铜绿微囊藻的去除率分别达到81.19%、91.41%、91.82%;4个处理组铜绿微囊藻的叶绿素a含量均显著低于对照组.添加稳定期滤液后,铜绿微囊藻MDA含量显著升高,SOD活性先升高后降低;在对光合色素的影响中,类胡萝卜素受到的影响不如叶绿素a显著.结果表明,枯草芽孢杆菌对铜绿微囊藻的抑制效果是通过分泌胞外物质实现的,且分泌物具有很强的热稳定性.推测该胞外分泌物能够破坏光合色素,影响光合作用,抑制藻细胞的生长;同时抑制SOD活性,使细胞膜脂过氧化程度不断加深,进而破坏藻细胞的完整性,表现出对藻很强的抑制效果.     

Isolation and identification of a bioflocculant-producing strain and optimisation of cultural conditions via a response surface model

A bioflocculant-producing strain named MY6-2 was isolated from a mixed activated sludge by a nitrogen-free medium. Based on 16S rDNA sequence, biochemical and morphological characteristics, the strain MY6-2 was identified as Bacillus mucilaginosus. The chemical analysis indicated that the flocculant MY6-2 was mainly composed of extracellular polysaccharide. The result of the composition of the medium showed that MY6-2 was able to generate bioflocculant in a nutrient-poor medium that consisted of 5?g?L^?1 sucrose and no nitrogen source. Response surface methodology (RSM) was used to optimise the fermentation condition of MY6-2 for maximum flocculating activity by using central composite design (CCD). Based on the result, the optimum conditions were as follows: 100?mL of broth in a 250?mL Erlenmeyer flask, initial pH 8.0 and inoculum concentration 10%, respectively. The highest flocculating rate of 90% was achieved under these conditions by adding 0.5?mL fermentation supernatant to 95?mL of Kaolin suspension.     

Microbiological evaluation of 4-(6-chloro-7H-purin-8-ylamino)-3-nitro-chromen-2-one with a comparison to antibacterial activity of standard drugs

Nucleophilic substitution of 4-chloro-3-nitro-2-oxo-2H-chromen-2-one 3 by 6-chloro-7H-purin-2-ylamine (a), yielded the corresponding 4-(6-chloro-7H-purin-8-ylamino)-3-nitro-chromen-2-one 4. The chemical structure of synthesized compound was characterized using IR and NMR spectra, and elemental analyses. The purified synthesized compound 4 was tested at concentrations 1, 3, 5 mg/ml for its antibacterial activity against three bacterial cultures: Staphylococcus aureus, Escherichia coli, and Bacillus cereus. The antibacterial activity of synthesized compound was compared to antibacterial activity of standard antibiotics cephalexine and streptomycin.     

城市居家环境空气细菌群落结构特征

在北京市5个方向(东南西北中)共选取31户有小孩的家庭于2009年11月至2010年10月研究了城市居家环境空气细菌的群落结构特征.结果表明,从分离的632株空气细菌中共鉴定出43属细菌,其中革兰氏阳性菌32属,革兰氏阴性菌11属.优势菌属依次为微球菌属(Micrococcus)、芽孢杆菌属(Bacillus)、葡萄球菌属(Staphylococcus)和库克氏菌属(Kocuria), 分别占25%~31%,12%~17%, 10%~15%,9%~18%,4属细菌百分比约占63%~71%.在北京市取样的31户家庭中,空气细菌浓度范围为47~12341cfu/m3,平均值为1821cfu/m3.总体上,春季和夏季空气细菌浓度分别为2967cfu/m3和1742cfu/m3,明显高于秋季和冬季的1334cfu/m3和1242cfu/m3(P<0.05).北京市居家环境空气细菌浓度男孩家庭(2123cfu/m3)明显高于女孩家庭(1511cfu/m3)(P<0.01).     

Biodegradation of Hexachlorocyclohexane by two species of Bacillus isolated from contaminated soil

Biodegradation of α, β, γ and δ hexachlorocyclohexane (HCH) isomers was studied in broth medium and soil microcosm by Bacillus circulans and Bacillus brevis isolated from contaminated soil. Degradation of α and γ isomers by both the bacterial isolates was higher than thermodynamically stable β and δ isomers. However, B. circulans was found more effective than B. brevis for β and δ isomers. Maximum rate of degradation was recorded at 150 mg/L followed by 100 and 50 mg/L. Soil microcosm study revealed maximum degradation of HCH isomers in the treatment containing natural soil, pesticide and bacterial inocula than the treatment having sterilised soil, pesticide and bacterial isolates. Chloride release was positively co-related with HCH degradation in broth medium as well as in soil microcosm, suggesting that B. circulans and B. brevis hold promising potential by having efficient enzyme(s) required for dechlorination of HCH from contaminated sites.